The overall objective of this project is to develop a validated fully automated, portable, point-of-care nucleic acid detection system for rapid diagnosis of hemorrhagic fever syndromes caused by NIAID category A-C biodefense pathogens. The laboratory diagnosis of viral hemorrhagic fever is currently complicated by the need for specialized laboratories and technical expertise. The resulting delays in definitive diagnosis of suspected viral hemorrhagic fever is a problem for clinical care in endemic regions and for biodefense readiness in developed countries. IQuum, Inc. has a proprietary "laboratory-in-a-tube" (Liat(tm)) technology that represents a promising approach to provide diagnostic capability for hemorrhagic fever viruses to sentinel or field laboratories. The Liat system, consisting of a proprietary assay tube system and analyzer device, provides a closed system for rapid, automated specimen handling and nucleic acid detection. This project proposes a cooperative research and development program between virologists at the University of Massachusetts, engineers at IQuum, Inc, and collaborators in tropical areas in the Americas to advance the development of the Liat system specifically for the rapid and sensitive detection of viral hemorrhagic fever agents. The Specific Aims of the project are: 1. To advance the engineering of the Liat Analyzer for improved reliability, ease of manufacture, and enhanced multiplex assay capability assays, thus yielding analyzers suitable for deployment in sentinel and field laboratories. 2. To define and optimize assays for diagnosis and prognostic evaluation of viral hemorrhagic fever syndromes caused by NIAID category A-C viruses. Our focus will be on development of assays for detection and quantitation of dengue viral RNA in serum and whole blood, and will extend to development of assays for prognostic markers in dengue and for detection of other hemorrhagic fever pathogens. 3. To develop a portfolio of pre-clinical and clinical data suitable for an IDE filing with the FDA for approval of the Liat differential diagnostic assay for dengue virus infection. This project will advance the development of a novel closed, fully automated assay platform suitable for front-line (point of care) detection of viruses that cause hemorrhagic fevers. Assay validation will focus initially on a commercially viable assay for detecting dengue virus and be extended to detection of other viruses. [unreadable] [unreadable] [unreadable]